The analysis of specimens for the presence or absence of target bacterial analytes, wherein the specimen is placed on a sterile growth media, is well known. Various bacteria can be detected in this manner in water, food stuffs; and in sample biological specimens, such as urine, cerebrospinal fluid, pleural fluid, ascites, joint fluid, stool, and the like. This testing procedure relies on the ability of the target analytes to replicate in the growth media to the extent that visible colonies of the target analytes can be observed. The various types of bacteria can be differentially labeled so as to aid in the differentiation between different bacterial colonies which grow in the media. When this analytic procedure is employed, the time necessary to form detectable colonies can be as short as one day, and as long as several weeks.
J. L. Burg et al describe a real time fluorescence detection procedure of RNA amplified by Q.beta. replicase, in Vol. 230 of Analytical Biochemistry (pp 263-272) 1995, Academic Press, Inc. The aforesaid detection procedure is non-specific and requires a considerable amount of sample preparation in order to function properly. Additionally, the Burg et al procedure cannot be used to detect DNA in a sample, and cannot detect more than one RNA target at a time. Quantification of any amplified RNA is a function of the time delay between the beginning of the procedure and the presence of detectable fluorescence in the sample container.
It would be highly desirable to be able to detect the presence or absence of one or more replicable target nucleotide sequence analytes in a single test, and quantify the target analytes in a short period of time. It would likewise be desirable to be able to detect organisms which cannot replicate in acellular systems, and to detect DNA genomes thought to be present in most life forms.